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NUTRACEUTICALS AND FUNCTIONAL FOODS, FDSC 6323, 2020

TAKE HOME EXAM III

Navam Hettiarachchy

Given on Thursday April 23, 2020 (Due date: Tuesday April 28 th, at 12:30 PM. Minus 15 points
per day will be deducted from the total score of 100 for late submission)

Please write your name and answer all the questions. Type one side of the paper only. A soft
copy of the answers to all the questions must be submitted as an attachment to my email
(nhettiar@uark.edu with a copy to Soma (Soma Mukherjee <soma.mukherjee75@gmail.com>;
Soma Mukherjee <sm113@uark.edu). Points for each question is give within () at the end of each
question. This is not a team exam and has to be answered independently. Please cut and paste
each question, and under each type in the answer.

Topics covered: 1. Nutraceutical Beverages, 2. Nutrigenomics, 3. Bioactive peptides as

functional food for health, 4. 3- D printing, 5. Biodegradable biocompatible nanoparticles as
carriers of active ingredients in nutraceuticals and healthcare, 6. General question covering the
entire syllabus

1.1 Name the bioactive phytochemicals/compounds of interest in cranberry and green tea
(3 points). Name their health benefits (3 points). Why are phenolic compounds
considered potent antioxidants? (2 points)

Bioactive compounds in cranberry are proanthocyanidins, reseveratrol, ellagic acid,

tocotrienols, lignans, etc. Bioactive compounds in green tea are epigallocathecin-3-
gallate, epicatechin-3-gallate, epigallocatechin, epicatechin, caffeine, gallic acid,
theobromine, theaflavin monogallate, theaflavin digallate, thearubigins, etc. Health
benefits of proanthocyanidins is preventing adhesion of certain types of E. coli
associated with urinary tract infections to uroepithelial cells and preventing adhesion
of H. pylori associated with stomach ulcer, and specific bacteria in mouth associated
with dental plaque and gingivitis. Proanthocyanidins also has benefits as antioxidants
and reducing total and LDL cholesterol. Health benefits of epigallocatechin-3-gallate are
promoting angiogenesis (growth capillary blood vessels) through Vascular Endothelial
Growth Factor (VEGF), scavenging free radicals that can damage DNA, regulating and
inhibiting cancer growth and killing cells, and decreasing metastasis (spread of a
disease from one organ or part to another non-adjacent organ). Theaflavins has
benefits such as halting metastasis, anticlastiogenic (causing disruption or breakages in
chromosomes). Thearubigins has benefits as an anticariogenic. Phenolic compounds
considered as potent antioxidants because phenolic hydroxyl groups act as good
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hydrogen donors (hydrogen-donating antioxidants can react with reactive oxygen and
reactive nitrogen species in a termination reaction) which leads to breaking the cycle of
generation of new radicals. The reaction will generate a radical form of the antioxidant
which possess a much greater chemical stability than the initial radical. The interaction
of the hydroxyl groups of phenolics with the -electrons of the benzene ring provides
the molecules special properties, especially the ability to generate free radicals where
the radical is stabilized by delocalization. The antioxidant capacity of phenolic
compounds is also correlated to their ability to chelate metal ions involved in the
production of free radicals.

1.2 “25 grams of soy protein reduce the risk of coronary heart disease”. Hence, there is
demand by the consumer for soy-based nutritional beverages. What are the challenges
in using soy in beverages? (3 points). How will you overcome these challenges? (2
points)

The challenges in using soy in beverages are correlated to the low content of calcium
that requires exogenous resources. Therefore, it will be challenging to ensure
bioavailability and homogeneous suspension of calcium in finished products. In order
to overcome these challenges, we can make calcium fortified soy beverages and
neutralized soy protein isolate (SPI) that showed excellent bioavailability.

1.2 How do divalent cations negatively affect protein solubility in a beverage? (2 points).
How will you overcome this negative effect? (2 points)

Divalent cations such as Ca, Mg, Zn can interact to the negatively charged COOH ends
of protein and cause protein aggregation. This interaction negatively affects protein
solubility in a beverage. In order to overcome this negative effect, we can add chelating
agents (such as citrates and phosphates) to make ions unavailable to interact with
protein.

1.3 Green tea extract and other polyphenolic compounds precipitate protein in beverages.
Innovatively discuss how this deleterious effect can be prevented (2 points)

In order to address this deleterious effect, we can suspend protein in beverages by

using gum stabilizers. Gums help build intermolecular bonding through electrostatic
linkages. This creates interstitial matrix making the protein movement in the fluid less
freely or by increasing the viscosity of the beverage to show gravity movement.
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2.1 What are the two general types of genome editing techniques? Provide an example for
each type (4 points).

There are the two general types of genome editing techniques, engineered nucleases
and recombination-based approaches. Example of engineered nucleases is
transcription activator-like effector nucleases (TALEN) and clustered regularly
interspaced short palindromic repeats (CRISPR). Example of recombination-based
approaches is recombinant adeno-associated virus (rAAV).

2.2 What are the practical applications of data generated by genomics? How will you apply
the data to integrate nutraceuticals and functional foods? (5 points)

Practical applications of data generated by genomics are:

- Comprehensive study of microbial pathogenesis and the interaction between

pathogens and their hosts.

- Identification of genes encoding systems that are unique to bacteria or a particular

pathogen.

- Discovery of microbial molecular markers associated with substantial variance in

the risk and severity of disease

- Oral plant vaccines, which use DNA and transgenes to create surface antigens that
stimulate immunity when consumed

- Selection of potential candidates for the rational development of new theraupetic

agents and vaccines

- Screening for thalassemia’s: tests have been evolved that use the polymerase chain
reaction (PCR) to observe the gene mutations that are responsible for creating the
structure of the hemoglobin molecule

- An application of genomics in the field of medicine that allows individualized

information about a patient’s genetic makeup to direct the type of treatment they
receive (precision medicine)

- Biotechnology applications include the creation of a partially synthetic species of

bacteria

In order to apply the data to integrate nutraceuticals and functional foods, we can
employ two strategies, the traditional genomics approach and the system biology
approach. The traditional genomics approach: specific genes and proteins, the
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expression of which is influenced by nutrients, are identified using genomic tools

such as transcriptomics, proteomics, and metabolomics which subsequently allows
the regulatory pathways through which diet influences homeostatis to be
identified. Transgenic mouse models and cellular models are essential tools. The
systems biology approach: gene, protein and metabolites signature that are
associated with specific nutrients, or nutritional regimes, are catalogued, and might
provide ‘early warning’ molecular biomarkers for nutrient-induced changes to
homeostatis.

2.3 Currently there is an increase in research interest on nutrigenomics and nutrigenetics

to cure chronic diseases. What would be your approach in using nutraceuticals as
dietary signals to add value to the ongoing research on obesity? (5 points)

My approach in using nutraceuticals as dietary signals to add value to the ongoing

research on obesity is by assessing the effect of certain nutraceuticals compound on
certain gene expression that can cause obesity. There are some nutraceuticals
compound that have been proven to control obesity. I will assess the effectivity of
these compounds in suppressing certain causative genetics factors that can contributes
to obesity. I will compare effectivity of two different nutraceuticals compounds to
suppress the expression of certain causative genetics factors that can contributes to
obesity in male and female population. Here I will use bioinformatics technologies such
as genomics (establish DNA sequence) and transcriptomics (patterns of gene
expression).

2.4 Provide two examples of short-term goals of nutrigenomics (4 points)

Short-term goals of nutrigenomics are:

- To identify the dietary signals

- To elucidate the dietary sensor mechanisms

3.1 Bioactive peptides can be released: a. in digestive system, b. during fermentation, from
microorganisms or plants, or c. combinations. These bioactive peptides have several
functionalities and applications. Outline a study that you propose to conduct to obtain
a pure peptide from rice bran and test for the inhibition of beta amyloid that
contributes to neurotoxicity in neuroblastoma cells. (7 points )

The outline of the study involves some steps: preparation, isolation, hydrolysis,
fractionation (UF), gastrointestinal resistance, and bioactivities testing. First, rice bran is
prepared, the protein and peptide hydrolysate are isolated from the rice bran. Protein
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is hydrolyzed using particular enzyme (e.g. trypsin) to obtain gastrointestinal juices

resistant peptides. Then, peptide hydrolysates are fractionated. Fractionated peptide
hydrolysates are subjected to purification (using HPLC), the amino acids are analyzed
through the MALDI profile and amino acid sequencing. After that, these peptide
hydrolysates are tested for the inhibition of neurotoxic β-amyloid peptide (1-42) in
neuroblastoma cells

3.2 Describe the mechanism of action of peptides as ACE-I inhibitor in the prevention of
hypertension (4 points)

Peptides as ACE-I inhibitor in the prevention of hypertension act by blocking

Angiotensin-I Converting Enzyme. Therefore, it will prevent the conversion (hydrolysis
by removing His-Leu from C-terminal) of angiotensin I (a decapeptide) to Angiotensin II,
octapeptide (a vasoconstrictor). This will directly influence blood pressure.

3.3 What is Miso? (2 points) Name the bioactives in miso that has anticancer activities (3
points)

Miso is a Japanese fermented soybean product. Bioactives compound in miso which

have anticancer activity are dietary fiber (preventing colon cancer), trypsin inhibitor,
isoflavones (such as genestein).

3.4 How do oligosaccharides lower the risk of colon cancer? (2 points)

Oligosaccharides promote bifidobacterial in the colon. High levels of bifidobacterial in

intestine associated with low risk of colon cancer.

4.1. What are the advantages and limitations of Inkjet printing in food 3- D printing
techniques? (4 points)

Advantages of inkjet printing in food 3-D printing are: excellent printing resolution and
relatively clean process. Whereas, limitations of inkjet printing in food 3-D printing are:
mostly for 2D surface filling and image decoration and limited choice of materials.

4.2 What is the future trend in 3-D printing for personalized food? (3 points)

The future trend in 3-D printing for personalized food is generating food for people
with certain diseases such as diabetes that requires careful monitoring on the blood
glucose level and also for the elderly population who has limitation in the digestive
function and to ensure that their nutritional needs is fulfilled. Besides that, the future
trend in 3-D printing for personalized food are not just related to the diseases and age
population but also to ensuring its sustainable availability. It means that the future
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trend in 3-D printing for personalized food will be related to the mass production not
only limited to small scale production. Besides that, the other future trend of this
technology for personalized food is the accessibility of this technology in order to
enabling people to generate and use it at home through the simpler application that
can be done by common people. The future development is also related to the
development of different print-heads that can be carried by the mobile printers for
printing food.

4.3 Name three specific innovative/unique ingredients that you will select for obesity
patients in producing a personalized food product using 3- D printing technology.
Justify your selection (5 points)

There are three specific ingredients that I will choose for obesity patients in producing
a personalized food product using 3-D printing technology. First, I will choose whole
grains-whole wheat because it will be digested more slowly than refined grains.
Therefore, they have a milder effect on blood sugar and insulin, which presumably
suppress hunger. The second ingredient will be vegetables and fruits. Fruit and
vegetables can prevent weight gain because of the presence of fiber that slows
digestion so that curbing hunger. The third ingredients will be seaweed because it has
high fiber content and also alginate in seaweed presumably increase satiety, reduce
energy intake and support weight reduction.

4.4 What are the three major barriers/limitations in commercializing 3-D printed food
products that were discussed in the class? (3 points)

The three major barriers/limitations in commercializing 3-D printed food products are
more research need to be conducted to ensure the quality of the food products
generating from 3-D printing because it is a new field, variability of the products (it will
be hard to maintain the similarity of the products because of the factors such as
machine, ingredients, printing processing), and the availability of the products in large
batch (the ability of 3-D printing to do mass production is still limited).

5.1 What is nanotechnology? (2 points)

Nanotechnology is science, engineering, and technology conducted at the nanoscale,

which is about 1 to 100 nanometers.

5.2 Is nanotechnology an independent technology like chemistry or biology? (2 points)

Why? (3 points)
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No, nanotechnology is not an independent technology like chemistry or biology

because it has an intricate interdisciplinary nature that combines the application of
material science, physics, and chemistry.

5.3 Give three examples for each: BD (Biodegradable) and BCs (Biocompatible)
nutraceuticals and healthcare (3 points)

Examples of BD (Biodegradable) are chitosan, gelatin, and polycaprolactone. Examples

of BCs (Biocompatible) are polylactic acid (PLA), poly lactic-co-glycolic acid (PLGA), and
polyethylene glycol (PEG).

5.4 Describe how you will engineer nanoparticles and load nutraceuticals for effective
delivery (4 points). How will you ensure that the encapsulated nutraceutical in
nanoparticles is stable and do not lose bioactivity? (4 points).

In order to engineer nanoparticles and load nutraceuticals for effective delivery, first it
is started with the NP core (inner material) which is not stable. Then the integration of
shell to coat the NP core will help with the stability issue (and also any other purposes
such as surface modification, the ability to increase the functionality, dispersibility, etc.)
but will make it not soluble in water. The core/shell type nanoparticles consist of a
different combination in close interaction, such as inorganic/inorganic,
inorganic/organic, organic/inorganic, etc. Then the core/shell is coated with polymer
and this polymer coating can be attached with biomolecule to have targeting effect. In
order to load nutraceuticals, there are two strategies that can be employed by
encapsulating nutraceuticals in BD BC NPs (50-150 nm NPs) and conjugation of the
nutraceuticals on the surface of BD BC NPs (50-150 NPs). In order to ensure stability of
the nutraceutical in nanoparticles, I will conduct some tests such as microbial
contamination after synthesis and stability to microbial attack. Microbial contamination
after synthesis is carried out on agar plates (for bacteria and yeast/molds) at 37℃ for
24-48 hours without preservatives and stability to microbial attack is conducted by
incubating at 0-4, 25, and 42-45℃ for 3-6 months. Another test that need to conduct is
related to the physical stability (pH, temperature, color and texture, mechanical phase
separation, and viscosity) and also chemical stability. Chemical stability study includes
smell or aroma and MS-based composition analysis over time: 3-6 months (samples
must be kept at various temperatures).

5.5 What are the advantages of bioactive delivery with BD, BC NPs? (NP-Nanoparticle) (3
points)

The advantages of bioactive delivery with BD, BC NPs are:

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- Enhancement of biological stability including solubility

- Improvement of cellular transport

- Release of the active component inside the cell

- Significantly greater bioavailability and bioactivity

- Site-specific targeting

- Longer half-life in the body

- Minimal side effects

- Enhanced therapeutic value

General Impact for your healthy life style

6. This class has provided you with simplified integrated approach of various
methods/techniques, approaches, and opportunities in enhancing your learning skills in
basic and advanced nutraceutical and functional foods. Describe how your approach in
food selections and practices is contributing to remain healthy (4 points). In your
opinion can immunomodulatory bioactive phytochemical/compounds when ingested
help recovery from COVID-19 better than without it? Why? (5 points)
My approach in food selection and practices to remain healthy is to choose food with
high fiber contents and also contain enough vitamins, minerals besides consuming food
supplements. My focus in high fiber contents is due to the beneficial effect of the food
with high fiber content on digestive system, weight control, and also preventive effect
on certain disease (such as colon cancer). Besides focus on food. I also balance the diet
with physical activity by doing exercise regularly to maintain physical and mental
health. The last point that is also important is getting enough sleep.
In my opinion ingestion of immunomodulatory bioactive phytochemical/compounds
presumably can help recovery from COVID-19 to some extents regarding to the severity
of the symptoms, another illness in combination with COVID-19 that may be contracted
by the patient, the other drugs that is consumed by the patient, effective formulation
for the patient with this condition. All of these factors need to be confirmed later with
the adequate research design. We need to consider to what extent that this compound
can effectively work to help patient to recovered from the COVID-19 (in which stage
that the patient needs to reach to be able to get the benefits from the
immunomodulatory bioactive phytochemical/compounds after the intubation). The
most important thing needs to be considered is the potential of interaction between
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the immunomodulatory bioactive compound with the drugs that are taken by the
patient, we need to know whether it will be harmful or not or diminish each other’s
effect. We need to carefully address this potential issue. Incorporating this
immunomodulatory compound into the proper formulation also needs to be
considered so that the patients can get benefit to recover from COVID-19.